Method and device for detecting a component in a sample

Abstract

The present disclosure provides a biosensor for detecting the presence of and/or the amount of at least one fungal plant pathogen in a sample, comprising: a support structure; at least two interdigitated electrodes coupled to the support structure, wherein at least one of the interdigitated electrodes is functionalized with a linker coupled to at least one biological component that recognizes the at least one fungal plant pathogen; and an impedance measurement circuit coupled to the at least two interdigitated electrodes. The present disclosure also provides methods of detecting the presence of and/or the amount of at least one fungal plant pathogen in a sample, methods of making the biosensor described herein, as well as methods and uses of using the herein described biosensor for detecting the presence of and/or amount of at least one fungal plant pathogen.

What is claimed is: 
     
         1 . A biosensor for detecting the presence of and/or the amount of at least one fungal plant pathogen in a sample, comprising:
 a support structure;   at least two interdigitated electrodes coupled to the support structure, wherein at least one of the at least two interdigitated electrodes is functionalized with a linker comprising a boronic acid functional group coupled to at least one antibody or fragment thereof that recognizes the at least one fungal plant pathogen; and   an impedance measurement circuit coupled to the at least two interdigitated electrodes,   wherein the at least one antibody or fragment thereof specifically binds at least one fungal plant pathogen that infects a canola plant, a wheat plant, a barley plant, a corn plant, a rice plant, a millet plant, a sorghum plant, or a combination thereof, or  Fusarium graminearum, F.avenaceum, F. poae, F.sporotrichioedes, Puccinia  spp.,  Puccinia triticina, P. recondite, P. striiformis, Erysiphe graminis  fsp.  tritici, Glomerella graminicola  (anamorph  Colletotrichum graminicola ),  Pyrenophora tritici - repentis  (telomorph) and  Drechslera tritici - repentis, Pyrenophora trichostoma, Urocystis agropyri, Sclerotinia borealis, Septoria  spp.,  Stagnospora  spp.  Parastagonospora nodorum, Pyrenophora  spp.  teres tares, Pyrenophora teres maculate, Claviceps purpurea, Alternaria  spp.,  Heminthosporium  spp,  Pseudocercosporella herpotrichoides, Glomerella graminicola  (anamorphic),  Colletotrichum graminicola, Fusarium verticillioides, Gibberella zeae, Aspergillus flavus, A. parasiticus, Lasiodiplodia theobromae, Physoderma maydis, Exserohilum turcicum, Cochliobolus heterostrophuspp, Cercospora zeae - maydis  and  Cercospora zeinaor, Cochliobolus carbonum, Stenocarpella maydis, Puccinia polysora, Magnaporthe oryzae, Cochliobolus miyabeanus, Ascochyta oryzae, Drechslera gigantean, Microdochium albescens, Cercospora oryzae, Puccinia graminis  f.sp.  oryzae, Uromyces coronatus, Ramularia oryzae, Bipolaris setariae, Cercospora penniseti, Curvularia penniseti, Dactuliophora elongata, Drechslera dematioidea, Claviceps fusiformis, Exserohilum rostratum, Beniowskia sphaeroidea, Myrothecium roridum, Phyllosticta penicillariae, Pyricularia grisea, Puccinia substriata, Moesziomyces penicillariae, Sclerotium rolfsii, Gleocercospora sorghi, Sarocladium strictum  (syns  Acremonium strictum ),  Cephalosporium acremonium, Macrophomina phaseolina, Claviceps africana, Sphacelia sorghi, Fusarium  spp.  moniliforme  (syn.  Gibberella fujikuroi ),  F. thapsinum  (syn.  G. thapsina ),  Aspergillus  spp.,  Fusarium andiyazi, F. nygamai, Penicillium  spp.,  Cercospora sorghi, Passalora fusimaculans  (syn.  Cercospora fusimaculans ),  Setosphaeria turcica  (syns.  Exserohilum turcicum, Helminthosporium  spp.  turcicum ),  Periconia circinata, Ramulispora sorghicola, Gibberella fujikuroi  (syns.  Fusarium moniliforme  var.  subglutinans G. fujikuroi  var.  subglutinans, G. intermedia, F. proliferatum ),  Ascochyta sorghi, Puccinia purpurea, Sclerotium rolfsii  (syn.  Athelia rolfsii ),  Ramulispora sorghi, Phyllachora sacchari, Bipolaris sorghicola  (syns.  B. cookei, Helminthosporium cookei. ),  Gloeocercospora sorghi,  or any combination thereof.   
     
     
         2 . The biosensor of  claim 1 , wherein the at least one antibody or fragment thereof specifically binds a spore of the at least one fungal plant pathogen or at least one airborne ascospore or conidiospore released from the apothecia. 
     
     
         3 . The biosensor of  claim 1 , wherein the linker is a SAM, a macromolecule, or a thin layer of polymer. 
     
     
         4 . The biosensor of  claim 3 , wherein the linker is a SAM that comprises a thiol head group. 
     
     
         5 . The biosensor of  claim 1 , wherein fingers of a pair of interdigitated electrodes, arranged together in an array, have an about square shape. 
     
     
         6 . The biosensor of  claim 1 , wherein the biosensor is a non-faradaic biosensor. 
     
     
         7 . The biosensor of  claim 1 , coupled to an ascospore trap. 
     
     
         8 . A method of making a biosensor for detecting the presence of and/or the amount of at least one fungal plant pathogen in a sample, comprising:
 contacting an interdigitated electrode with an aqueous solution comprising a linker, the interdigitated electrode being in electrical communication with at least a second interdigitated electrode;   contacting the interdigitated electrode with an aqueous solution comprising EDC and NHS;   contacting the interdigitated electrode with an aqueous solution comprising APBA; and   contacting the interdigitated electrode with a solution comprising at least one antibody or fragment thereof that specifically binds at least one fungal plant pathogen that infects a canola plant, a wheat plant, a barley plant, a corn plant, a rice plant, a millet plant, a sorghum plant, or a combination thereof, or  Fusarium graminearum, F.avenaceum, F. poae, F. sporotrichioedes, Puccinia  spp.,  Puccinia triticina, P. recondite, P. striiformis, Erysiphe graminis  fsp.  tritici, Glomerella graminicola  (anamorph  Colletotrichum graminicola ),  Pyrenophora tritici - repentis  (telomorph) and  Drechslera tritici - repentis, Pyrenophora trichostoma, Urocystis agropyri, Sclerotinia borealis, Septoria  spp.,  Stagnospora  spp.  Parastagonospora nodorum, Pyrenophora  spp.  teres tares, Pyrenophora teres maculate, Claviceps purpurea, Alternaria  spp.,  Heminthosporium  spp,  Pseudocercosporella herpotrichoides, Glomerella graminicola  (anamorphic),  Colletotrichum graminicola, Fusarium verticillioides, Gibberella zeae, Aspergillus flavus, A. parasiticus, Lasiodiplodia theobromae, Physoderma maydis, Exserohilum turcicum, Cochliobolus heterostrophuspp, Cercospora zeae - maydis  and  Cercospora zeinaor, Cochliobolus carbonum, Stenocarpella maydis, Puccinia polysora, Magnaporthe oryzae, Cochliobolus miyabeanus, Ascochyta oryzae, Drechslera gigantean, Microdochium albescens, Cercospora oryzae, Puccinia graminis  f.sp. oryzae,  Uromyces coronatus, Ramularia oryzae, Bipolaris setariae, Cercospora penniseti, Curvularia penniseti, Dactuliophora elongata, Drechslera dematioidea, Claviceps fusiformis, Exserohilum rostratum, Beniowskia sphaeroidea, Myrothecium roridum, Phyllosticta penicillariae, Pyricularia grisea, Puccinia substriata, Moesziomyces penicillariae, Sclerotium rolfsii, Gleocercospora sorghi, Sarocladium strictum  (syns Acremonium strictum),  Cephalosporium acremonium, Macrophomina phaseolina, Claviceps africana, Sphacelia sorghi, Fusarium  spp.  moniliforme  (syn.  Gibberella fujikuroi ),  F. thapsinum  (syn.  G. thapsina ),  Aspergillus  spp.,  Fusarium andiyazi, F. nygamai, Penicillium  spp.,  Cercospora sorghi, Passalora fusimaculans  (syn.  Cercospora fusimaculans ),  Setosphaeria turcica  (syns.  Exserohilum turcicum, Helminthosporium  spp.  turcicum ),  Periconia circinata, Ramulispora sorghicola, Gibberella fujikuroi  (syns.  Fusarium moniliforme  var.  subglutinans G. fujikuroi  var.  subglutinans, G. intermedia, F. proliferatum ),  Ascochyta sorghi, Puccinia purpurea, Sclerotium rolfsii  (syn.  Athelia rolfsii ),  Ramulispora sorghi, Phyllachora sacchari, Bipolaris sorghicola  (syns.  B. cookei, Helminthosporium cookei. ),  Gloeocercospora sorghi,  or any combination thereof.   
     
     
         9 . The method of  claim 8 , wherein the at least one antibody or fragment thereof specifically binds a spore of the at least one fungal plant pathogen or at least one airborne ascospore or conidiospore released from the apothecia. 
     
     
         10 . The method of  claim 8 , further comprising the step of: applying a PDMS mask to the interdigitated electrode. 
     
     
         11 . The method of  claim 8 , wherein the linker is a SAM that comprises a thiol head group and optionally, a boron functional group, a macromolecule, a thin layer of polymer, or 6-MHA and 11-MUA. 
     
     
         12 . The method of  claim 8 , wherein the biosensor is the biosensor of  claim 1 . 
     
     
         13 . A method of detecting the presence of and/or the amount of at least one fungal plant pathogen in a sample, comprising:
 measuring the impedance of a biosensor coupled to at least one antibody or fragment thereof that specifically binds at least one fungal plant pathogen that infects a canola plant, a wheat plant, a barley plant, a corn plant, a rice plant, a millet plant, a sorghum plant, or a combination thereof, or  Fusarium graminearum, F.avenaceum, F. poae, F.sporotrichioedes, Puccinia  spp.,  Puccinia triticina, P. recondite, P. striiformis, Erysiphe graminis  f.sp.  tritici, Glomerella graminicola  (anamorph  Colletotrichum graminicola ),  Pyrenophora tritici - repentis  (telomorph) and  Drechslera tritici - repentis, Pyrenophora trichostoma, Urocystis agropyri, Sclerotinia borealis, Septoria  spp.,  Stagnospora  spp.  Parastagonospora nodorum, Pyrenophora  spp.  teres tares, Pyrenophora teres maculate, Claviceps purpurea, Alternaria  spp.,  Heminthosporium  spp,  Pseudocercosporella herpotrichoides, Glomerella graminicola  ( anamorphic ),  Colletotrichum graminicola, Fusarium verticillioides, Gibberella zeae, Aspergillus flavus, A. parasiticus, Lasiodiplodia theobromae, Physoderma maydis, Exserohilum turcicum, Cochliobolus heterostrophuspp, Cercospora zeae - maydis  and  Cercospora zeinaor, Cochliobolus carbonum, Stenocarpella maydis, Puccinia polysora, Magnaporthe oryzae, Cochliobolus miyabeanus, Ascochyta oryzae, Drechslera gigantean, Microdochium albescens, Cercospora oryzae, Puccinia graminis  f.sp. oryzae,  Uromyces coronatus, Ramularia oryzae, Bipolaris setariae, Cercospora penniseti, Curvularia penniseti, Dactuliophora elongata, Drechslera dematioidea, Claviceps fusiformis, Exserohilum rostratum, Beniowskia sphaeroidea, Myrothecium roridum, Phyllosticta penicillariae, Pyricularia grisea, Puccinia substriata, Moesziomyces penicillariae, Sclerotium rolfsii, Gleocercospora sorghi, Sarocladium strictum  (syns  Acremonium strictum ),  Cephalosporium acremonium, Macrophomina phaseolina, Claviceps africana, Sphacelia sorghi, Fusarium  spp.  moniliforme  (syn.  Gibberella fujikuroi ),  F. thapsinum  (syn.  G. thapsina ),  Aspergillus  spp.,  Fusarium andiyazi, F. nygamai, Penicillium  spp.,  Cercospora sorghi, Passalora fusimaculans  (syn.  Cercospora fusimaculans ),  Setosphaeria turcica  (syns.  Exserohilum turcicum, Helminthosporium  spp.  turcicum ),  Periconia circinata, Ramulispora sorghicola, Gibberella fujikuroi  (syns.  Fusarium moniliforme  var.  subglutinans G. fujikuroi  var.  subglutinans, G. intermedia, F. proliferatum ),  Ascochyta sorghi, Puccinia purpurea, Sclerotium rolfsii  (syn.  Athelia Ramulispora sorghi, Phyllachora sacchari, Bipolaris sorghicola  (syns.  B. cookei, Helminthosporium cookei. ),  Gloeocercospora sorghi,  or any combination thereof;   contacting the sample with the biosensor;   measuring the impedance of the biosensor in contact with the sample; and   correlating the change of impedance with the presence and/or the amount of the at least one fungal plant pathogen in the sample.   
     
     
         14 . The method of  claim 13 , wherein the at least one antibody or fragment thereof specifically binds a spore of the at least one fungal plant pathogen or at least one airborne ascospore or conidiospore released from the apothecia. 
     
     
         15 . The method of  claim 13 , wherein the correlating step comprises correlating the change of impedance with a standard curve of known impedances of known amounts of the at least one fungal plant pathogen. 
     
     
         16 . The method of  claim 13 , wherein the impedances are measured by applying about 10 mV sinusoidal excitation perturbation at 0 V DC in a frequency range from about 10 Hz to about 1 MHz. 
     
     
         17 . The method of  claim 13 , wherein the at least one fungal plant pathogen is the apothecia of the at least one fungal plant pathogen or at least one airborne ascospore or conidiospore released from the apothecia. 
     
     
         18 . The method of  claim 13 , wherein the sample is captured in a high throughput ascospore trap. 
     
     
         19 . The method of  claim 13 , wherein the biosensor is the biosensor of  claim 1 .